Individual Phosphatidylcholine Species Analysis by RP-HPLC-ELSD for
Determination of Polyenylphosphatidylcholine in Lecithins
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Abstract
Polyenylphosphatidylcholine
(PPC), a subgroup of the bioactive agents in phosphatidylcholine (PC),
has been indicated to possess liver-protective effects. This study
aimed to investigate a promising and feasible method to determine
PC molecular species with a reverse phase (RP) high-performance liquid
chromatograph (HPLC) equipped with an evaporative light scattering
detector (ELSD). Chromatography was achieved using a C30 column and
an isocratic mobile phase consisting of acetonitrile/methanol/triethylamine
(40/58/2, v/v/v) at a flow rate of 1 mL/min, and ELSD detection was
performed using 80 °C for the drift tube and an air flow rate
of 1.8 L/min. To identify individual peaks on the chromatogram, MALDI-TOF-MS
was employed for initial detection, and then the results were used
to investigate the relationship between the retention time and fatty
acyl chains of each PC molecule. A linear correlation was observed
between the retention time and theoretical carbon number (TCN) of
individual PC species. The compositions of PC molecular species in
soybean and sunflower lecithins were similar to each other, and the
major PC molecular species were 1,2-dilinoleoyl-<i>sn</i>-glycero-3-phosphocholine (LLPC), 1-oleoyl-2-linoleoyl-<i>sn</i>-glycero-3-phosphocholine (OLPC), and 1-palmitoyl-2-linoleoyl-<i>sn</i>-glycero-3-phosphocholine (PLPC). The contents of LLPC
in soybean PC and sunflower PC were 40.6% and 64.3%, respectively